Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1991 Jan 15;273(Pt 2):311–316. doi: 10.1042/bj2730311

Identification of valine/leucine/isoleucine and threonine/alanine/glycine proton-spin systems of Escherichia coli adenylate kinase by selective deuteration and selective protonation.

I Bock-Möbius 1, M Brune 1, A Wittinghofer 1, H Zimmermann 1, R Leberman 1, M T Dauvergne 1, S Zimmermann 1, B Brandmeier 1, P Rösch 1
PMCID: PMC1150215  PMID: 1991031

Abstract

Adenylate kinase from two types of Escherichia coli strains, a wild-type and a leucine-auxotrophic strain, was purified. On the one hand, growing the leucine-auxotrophic bacteria on a medium containing deuterated leucine yielded E. coli adenylate kinase with all leucine residues deuterated. On the other hand, by growing the wild-type bacteria on deuterated medium with phenylalanine, threonine and isoleucine present as protonated specimens, 80% randomly deuterated enzyme with protonated phenylalanine, threonine and isoleucine residues could be prepared. Use of these proteins enabled identification of the spin systems of these amino acid residues in the n.m.r. spectra of the protein.

Full text

PDF
311

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bock I., Reinstein J., Brune M., Wittinghofer A., Rösch P. Proton-nuclear magnetic resonance studies of the aromatic spin systems of Escherichia coli adenylate kinase. J Mol Biol. 1988 Apr 20;200(4):745–748. doi: 10.1016/0022-2836(88)90486-x. [DOI] [PubMed] [Google Scholar]
  2. Brune M., Schumann R., Wittinghofer F. Cloning and sequencing of the adenylate kinase gene (adk) of Escherichia coli. Nucleic Acids Res. 1985 Oct 11;13(19):7139–7151. doi: 10.1093/nar/13.19.7139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bârzu O., Michelson S. Simple and fast purification of Escherichia coli adenylate kinase. FEBS Lett. 1983 Mar 21;153(2):280–284. doi: 10.1016/0014-5793(83)80624-3. [DOI] [PubMed] [Google Scholar]
  4. Ehresmann B., Imbault P., Weil J. H. Spectrophotometric determination of protein concentration in cell extracts containing tRNA's and rRNA's. Anal Biochem. 1973 Aug;54(2):454–463. doi: 10.1016/0003-2697(73)90374-6. [DOI] [PubMed] [Google Scholar]
  5. Haase G. H., Brune M., Reinstein J., Pai E. F., Pingoud A., Wittinghofer A. Adenylate kinases from thermosensitive Escherichia coli strains. J Mol Biol. 1989 May 5;207(1):151–162. doi: 10.1016/0022-2836(89)90446-4. [DOI] [PubMed] [Google Scholar]
  6. Lienhard G. E., Secemski I. I. P 1 ,P 5 -Di(adenosine-5')pentaphosphate, a potent multisubstrate inhibitor of adenylate kinase. J Biol Chem. 1973 Feb 10;248(3):1121–1123. [PubMed] [Google Scholar]
  7. Marion D., Wüthrich K. Application of phase sensitive two-dimensional correlated spectroscopy (COSY) for measurements of 1H-1H spin-spin coupling constants in proteins. Biochem Biophys Res Commun. 1983 Jun 29;113(3):967–974. doi: 10.1016/0006-291x(83)91093-8. [DOI] [PubMed] [Google Scholar]
  8. Müller C. W., Schulz G. E. Structure of the complex of adenylate kinase from Escherichia coli with the inhibitor P1,P5-di(adenosine-5'-)pentaphosphate. J Mol Biol. 1988 Aug 20;202(4):909–912. doi: 10.1016/0022-2836(88)90567-0. [DOI] [PubMed] [Google Scholar]
  9. Rance M., Sørensen O. W., Bodenhausen G., Wagner G., Ernst R. R., Wüthrich K. Improved spectral resolution in cosy 1H NMR spectra of proteins via double quantum filtering. Biochem Biophys Res Commun. 1983 Dec 16;117(2):479–485. doi: 10.1016/0006-291x(83)91225-1. [DOI] [PubMed] [Google Scholar]
  10. Reinstein J., Brune M., Wittinghofer A. Mutations in the nucleotide binding loop of adenylate kinase of Escherichia coli. Biochemistry. 1988 Jun 28;27(13):4712–4720. doi: 10.1021/bi00413a020. [DOI] [PubMed] [Google Scholar]
  11. Searle M. S., Hammond S. J., Birdsall B., Roberts G. C., Feeney J., King R. W., Griffiths D. V. Identification of the 1H resonances of valine and leucine residues in dihydrofolate reductase by using a combination of selective deuteration and two-dimensional correlation spectroscopy. FEBS Lett. 1986 Jan 1;194(1):165–170. doi: 10.1016/0014-5793(86)80070-9. [DOI] [PubMed] [Google Scholar]
  12. Stella V. J. Kinetics of synthesis of d 4 -succinic anhydride from succinic anhydride and d 1 -acetic acid. J Pharm Sci. 1973 Apr;62(4):634–637. doi: 10.1002/jps.2600620420. [DOI] [PubMed] [Google Scholar]
  13. Vetter I. R., Reinstein J., Rösch P. Complexes of Escherichia coli adenylate kinase and nucleotides: 1H NMR studies of the nucleotide sites in solution. Biochemistry. 1990 Aug 14;29(32):7459–7467. doi: 10.1021/bi00484a015. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES